title: Comparison of (bio-)transformation methods for the generation of metabolite-like compound libraries of p38α MAP kinase inhibitors using high-resolution screening.
abstract: Four hydrophobic p38α mitogen-activated protein kinase inhibitors were refluxed with 7.5% hydrogen peroxide at 80°C and irradiated with visible light in order to generate more hydrophilic conversion products. The resulting mixtures were analyzed in a high-resolution screening (HRS) platform, featuring liquid chromatographic separation coupled in parallel with a fluorescence enhancement based continuous-flow affinity bioassay towards the p38α mitogen-activated protein kinase and with high-resolution (tandem) mass spectrometry on an ion-trap-time-of-flight hybrid instrument. The results were compared with similar data where chemical diversity was achieved by means of electrochemical conversion or incubation with either human liver microsomes or cytochrome P450s from Bacillus megaterium (BM3s). In total, more than 50 conversion products were identified. The metabolite-like compound libraries studied are discussed in terms of the reactions enabled, the retention of affinity, and the change in hydrophilicity by modification, in summary the ability to generate bioactive, more hydrophilic potential lead compounds. In this context, HRS is demonstrated to be an effective tool as it reduces the effort directed towards laborious synthesis and purification schemes.
authors.affiliation: AIMMS Division of BioMolecular Analysis, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
authors.forename: David
authors.initials: D
authors.lastname: Falck
authors.forename: Fatie
authors.initials: F
authors.lastname: Rahimi Pirkolachachi
authors.forename: Martin
authors.initials: M
authors.lastname: Giera
authors.forename: Maarten
authors.initials: M
authors.lastname: Honing
authors.forename: Jeroen
authors.initials: J
authors.lastname: Kool
authors.forename: Wilfried M A
authors.initials: WM
authors.lastname: Niessen
mesh_terms: D001410:Bacillus megaterium / Q000378:metabolism; D001549:Benzamides / Q000737:chemistry; D002626:Chemistry, Pharmaceutical; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry; D057166:High-Throughput Screening Assays; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D007093:Imidazoles / Q000737:chemistry; D007211:Indoles / Q000737:chemistry; D008027:Light; D008099:Liver / Q000201:enzymology; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D048308:Mitogen-Activated Protein Kinase 14 / Q000037:antagonists & inhibitors*; D009281:Naphthalenes / Q000737:chemistry; D010100:Oxygen / Q000737:chemistry; D010777:Photochemistry; D010879:Piperazines / Q000737:chemistry; D047428:Protein Kinase Inhibitors / Q000737:chemistry; D011720:Pyrazoles / Q000737:chemistry; D011725:Pyridines / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D013329:Structure-Activity Relationship; D013844:Thiazoles / Q000737:chemistry; D010671:Phenylurea Compounds
chemical_list: D001549:Benzamides; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide; D007093:Imidazoles; D007211:Indoles; D048308:Mitogen-Activated Protein Kinase 14; D009281:Naphthalenes; D010100:Oxygen; D010879:Piperazines; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011725:Pyridines; D011994:Recombinant Proteins; D013844:Thiazoles; C000589975:1-(6-chloro-5-(4-(4-fluorobenzyl)-2,5-dimethylpiperazine-1-carbonyl)-3aH-indol-3-yl)-2-morpholinoethane-1,2-dione; C505166:N-(4-(2-ethyl-4-(3-methylphenyl)-1,3-thiazol-5-yl)-2-pyridyl)benzamide; C452139:doramapimod; D010671:Phenylurea Compounds; C093642:SB 203580
keywords: Chemical oxidation; High-resolution screening; Photochemistry; Physicochemical properties; p38α mitogen-activated protein kinase inhibitors